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[在中国首次从蚊子中分离出广平样病毒]

[First isolation of quang binh-like virus from mosquitoes in China].

作者信息

Feng Yun, Li Hong-Bin, Zhu Jin, Zhang Yu-Zhen, Yang Wei-Hong, Fan Jian-Hua, Liang Guo-Dong, Zhang Hai-Lin

出版信息

Bing Du Xue Bao. 2014 Jan;30(1):57-61.

Abstract

This study aims to investigate the distribution patterns of mosquito-borne viruses in Menghai County, Xishuangbanna Prefecture, Yunnan Province, China and to provide evidence for the prevention and control of mosquito-borne diseases. Mosquito samples were collected using mosquito lamps. Viruses were isolated from the samples by cell culture, and the isolates were identified by RT-PCR. The genomes of isolates were sequenced for phylogenetic analysis. In July 2012, a total of 1468 mosquitoes were captured in Daluo Town of Menghai County; they were divided into 32 pools, including Culex tritaeniorhynchus (28 pools, 1383 mosquitoes), Culex quinquefasciatus (2 pools, 66 mosquitoes), and Anopheles (2 pools, 19 mosquitoes). Golden hamster kidney cells (BHK-21) and Aedes albopictus cells (C6/36) were used for virus isolation. The results showed that C6/36 cells were susceptible to two isolates recovered from Culex tritaeniorhynchus (BNDL1205 and BNDL1227), with marked cytopathic effect (CPE) of cell fusion. By contrast, the two isolates could not cause CPE in BHK-21 cells. RT-PCR was performed for the two isolates using the flavivirus-specific primers FU2/cFD3, and a 800-bp amplicon was obtained from both of them. Phylogenetic analysis showed that the two isolates shared the same evolutionary branch with the Quang Binh virus (QBV) strain VN180, which had been isolated from Vietnam, with nucleotide sequence homologies of 83.4% and 82.9%, respectively. However, there existed relatively large differences in nucleotide sequence between them and other Culex flavivirus strains previously isolated in China and other regions. In light of the similarity between the two isolates and QBV, BNDL1205 and BNDL122 were referred to as Quang Binh-like virus, which were first reported in China.

摘要

本研究旨在调查中国云南省西双版纳州勐海县蚊媒病毒的分布模式,为蚊媒疾病的预防和控制提供依据。使用诱蚊灯收集蚊子样本。通过细胞培养从样本中分离病毒,并通过逆转录聚合酶链反应(RT-PCR)鉴定分离株。对分离株的基因组进行测序以进行系统发育分析。2012年7月,在勐海县打洛镇共捕获1468只蚊子;它们被分为32组,包括三带喙库蚊(28组,1383只蚊子)、致倦库蚊(2组,66只蚊子)和按蚊(2组,19只蚊子)。使用金黄仓鼠肾细胞(BHK-21)和白纹伊蚊细胞(C6/36)进行病毒分离。结果表明,C6/36细胞对从三带喙库蚊分离出的两个分离株(BNDL1205和BNDL1227)敏感,具有明显的细胞融合细胞病变效应(CPE)。相比之下,这两个分离株在BHK-21细胞中未引起CPE。使用黄病毒特异性引物FU2/cFD3对这两个分离株进行RT-PCR,从它们两者中均获得了一个800碱基对的扩增子。系统发育分析表明,这两个分离株与从越南分离出的广平病毒(QBV)株VN180共享同一进化分支,核苷酸序列同源性分别为83.4%和82.9%。然而,它们与先前在中国和其他地区分离的其他库蚊黄病毒株在核苷酸序列上存在较大差异。鉴于这两个分离株与QBV的相似性,BNDL1205和BNDL122被称为类广平病毒,这是在中国首次报道。

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